WO2020130626A1 - Aptamère d'adn se liant spécifiquement à ediii et utilisation correspondante - Google Patents

Aptamère d'adn se liant spécifiquement à ediii et utilisation correspondante Download PDF

Info

Publication number
WO2020130626A1
WO2020130626A1 PCT/KR2019/017998 KR2019017998W WO2020130626A1 WO 2020130626 A1 WO2020130626 A1 WO 2020130626A1 KR 2019017998 W KR2019017998 W KR 2019017998W WO 2020130626 A1 WO2020130626 A1 WO 2020130626A1
Authority
WO
WIPO (PCT)
Prior art keywords
ediii
dna aptamer
dengue virus
aptamer
dengue
Prior art date
Application number
PCT/KR2019/017998
Other languages
English (en)
Korean (ko)
Inventor
김윤근
반창일
Original Assignee
주식회사 엠디헬스케어
포항공과대학교 산학협력단
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 주식회사 엠디헬스케어, 포항공과대학교 산학협력단 filed Critical 주식회사 엠디헬스케어
Priority to CN201980084587.XA priority Critical patent/CN113227378B/zh
Publication of WO2020130626A1 publication Critical patent/WO2020130626A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/16—Aptamers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/08—RNA viruses
    • G01N2333/18—Togaviridae; Flaviviridae
    • G01N2333/183—Flaviviridae, e.g. pestivirus, mucosal disease virus, bovine viral diarrhoea virus, classical swine fever virus (hog cholera virus) or border disease virus
    • G01N2333/185—Flaviviruses or Group B arboviruses, e.g. yellow fever virus, japanese encephalitis, tick-borne encephalitis, dengue
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to a DNA aptamer specifically binding to dengue virus EDIII (Dengue virus envelope protein domain III; DENV EDIII), a composition for diagnosis of dengue fever comprising the same, and a dengue diagnosis kit.
  • dengue virus EDIII Dengue virus envelope protein domain III
  • DENV EDIII Dengue virus envelope protein domain III
  • Dengue fever is a severe flu-like condition.
  • the dengue virus belongs to a flavivirus, such as the yellow fever virus, and shows structural and pathological similarities. Symptoms include fever, severe headache, muscle and joint pain, and lymph node inflammation. Dengue fever itself has a low mortality rate, but there is a risk of complications such as hypothermia or hemorrhagic vomiting.
  • dengue virus EDIII (Dengue virus envelope protein domain III; DENV EDIII) is a domain III part located on the surface protein of dengue virus, and the flavivirus to which the dengue virus belongs is inside the capsid protein.
  • Genomic RNA genomic RNA
  • E envelope
  • M membrane protein
  • the envelope protein located on the outermost side of the flavivirus antigen is composed of domains I, II, and III, of which domain III is a location where viral receptors bind, and has attracted attention as a major target for diagnosis and immune research using serum. have.
  • Domain III is highly antigenic and has a linear epitope recognized by a virus-neutralizing single antibody. Diseases of the same flavivirus have a strong antigen-cross reaction between viruses, making it difficult for serological diagnosis. For example, there are reports that most infections have been misdiagnosed by a cross reaction between type 3 dengue fever, yellow fever, and St. Louis encephalitis. In order to solve this problem, the sequence of domain III having distinct characteristics even in viruses of the same genus is receiving attention (Registration No. 10-1520084, Republic of Korea).
  • aptamer has a very high affinity for a specific material and is stable, and has been developed a lot in recent years, and its application as a therapeutic and diagnostic sensor using it is actively progressing. Synthesis of aptamers is possible in a relatively simple way, and even cells, proteins, and even small organic substances can be targeted, enabling the development of new detection methods using them, and their specificity and stability are very high compared to antibodies already developed. Therefore, it is possible to develop therapeutic agents, drug delivery systems, and applications as diagnostic biosensors.
  • the present invention was devised to solve the above problems, and the specific binding capacity of the DNA aptamer prepared in the present invention to the dengue virus envelope protein domain III (DENV EDIII) was confirmed and based on this The invention was completed.
  • the object of the present invention is a DNA aptamer that specifically binds to the Dengue virus EDIII (Dengue virus envelope protein domain III; DENV EDIII), wherein the DNA aptamer comprises a nucleotide sequence of SEQ ID NO: 4, DNA To provide aptamers.
  • DENV EDIII Dengue virus envelope protein domain III
  • another object of the present invention is to provide a dengue fever diagnostic kit or a composition for dengue fever diagnosis, which includes a DNA aptamer.
  • the present invention is a DNA aptamer specifically binding to dengue virus EDIII (Dengue virus envelope protein domain III; DENV EDIII), wherein the DNA aptamer is the base sequence of SEQ ID NO: 4 It provides a DNA aptamer, characterized in that consisting of.
  • DENV EDIII Dengue virus envelope protein domain III
  • the DENV EDIII may be a domain protein included on the surface of the dengue virus antigen.
  • the present invention provides a composition for diagnosing dengue fever, which includes a DNA aptamer.
  • the present invention provides a dengue fever diagnostic kit comprising a DNA aptamer.
  • the present inventors have conducted extensive research to develop a novel biomarker for the diagnosis of Dengue fever, and DNA aptamer (DNA) with specific binding ability to Dengue virus envelope protein domain III (DENV EDIII) Aptamer) has been confirmed to have strong binding strength and excellent specificity with DENV EDIII, and thus it can be expected to have superior stability than the ELISA method using an existing antibody, and thus can be usefully used in the development of dengue fever diagnostic composition, dengue fever diagnostic kit, etc. It is expected to be.
  • DNA DNA aptamer
  • DENV EDIII Dengue virus envelope protein domain III
  • Figure 1 is a graph showing the results of confirming the binding degree (%) of ssDNA (single strand DNA) and DENV EDIII, and the screening process.
  • Figure 2 shows the result of confirming the Kd value for the DV4 aptamer sequence through fluorescence measurement.
  • Figure 3 shows the results of confirming whether the binding of DV1, DV2, DV3, DV4 aptamer with DENV EDIII protein through EMSA (Electrophoric Mobility Shift Assay) method.
  • the present inventors have conducted extensive research to develop a novel biomarker for the diagnosis of Dengue fever, and DNA aptamer (DNA) with specific binding ability to Dengue virus envelope protein domain III (DENV EDIII) Aptamer) confirmed that it has a strong binding force with DENV EDIII and excellent specificity, and completed the present invention.
  • DNA DNA aptamer
  • DENV EDIII Dengue virus envelope protein domain III
  • the present invention provides a DNA aptamer that specifically binds to DENV EDIII
  • Dengue virus EDIII (Dengue virus envelope protein domain III; DENV EDIII)
  • DENV EDIII domain III portion located on the surface protein of the Dengue virus, the Flavivirus to which the Dengue virus belongs
  • Genomic RNA is located inside the capsid protein, and the outer surface has a structure surrounded by dimers and membrane proteins (M) proteins.
  • M dimers and membrane proteins
  • the envelope protein located on the outermost side of the flavivirus antigen is composed of domains I, II, and III, of which domain III is a location where viral receptors bind, and is receiving attention as a major target for diagnosis and immune research using serum. have.
  • Domain III is highly antigenic and has a linear epitope recognized by a virus-neutralizing single antibody.
  • aptamer is a single-stranded DNA (ssDNA) or RNA having high specificity and affinity for a specific substance.
  • ssDNA single-stranded DNA
  • RNA RNA having high specificity and affinity for a specific substance.
  • Conventional methods using antibodies are relatively time-consuming and expensive because they are made using the immune system of a living body, and their stability may be problematic because they are proteins, whereas aptamers are relatively synthetic in synthesis. Since it is possible by a simple method and can target cells, proteins, and even small organic substances, it is possible to develop new detection methods using this, and the specificity and stability are very high compared to the antibodies already developed.
  • DNA aptamers were used for the specific detection of EDIII. Any DNA (ssDNA) or RNA capable of specific detection of DENV EDIII may correspond to the aptamer of the present invention, and may be preferably composed of the nucleotide sequence of SEQ ID NO: 4, but is not limited thereto.
  • the present invention provides a composition for diagnosing dengue fever, comprising the DNA aptamer.
  • composition of the present invention may further include a pharmacologically or physiologically acceptable carrier, excipient, and diluent in addition to the DNA aptamer.
  • a pharmacologically or physiologically acceptable carrier such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions according to a conventional method.
  • Carriers, excipients, and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl Cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, and mineral oil.
  • diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, and the like.
  • the present invention provides a dengue fever diagnostic kit comprising the DNA aptamer.
  • the kit includes the aptamer of the present invention that specifically binds to DENV EDIII, and the aptamer may be attached with detection labels widely used in the art, for example, biotin residues.
  • detection labels widely used in the art, for example, biotin residues.
  • the labeling material After introducing the labeling material to the biotin-attached aptamer, it can be used for detection, quantification and diagnosis of DENV EDIII through its analysis.
  • various known analytical labeling materials can be used, for example, fluorescence analysis using streptavidin, avidin, Cy3, Cy5, Alexa, BODIPY, Rhodamine or Q-dot, etc. Can.
  • aptamers for DENV EDIII detection can be labeled with common labeling substances such as other fluorescent substances, magnetic substances, dyes, enzymes, radioactive isotopes, and common detection means, for example, fluorescence. It can be detected through a microscope, Radioimmnodetection (RAID), or the like.
  • common labeling substances such as other fluorescent substances, magnetic substances, dyes, enzymes, radioactive isotopes, and common detection means, for example, fluorescence. It can be detected through a microscope, Radioimmnodetection (RAID), or the like.
  • kit is a variety of tools or reagents that can be used to qualitatively or quantitatively determine whether aptamer is bound to DENV EDIII in a sample, a support (substrate), buffer solution, reaction stopper, solubilizer, detergent or stabilizer It may further include.
  • the aptamer-fixed substrate may be, for example, a solid substrate, selected from the group consisting of polymers, glass, gold, paper, and biomembrane. More specifically, polystyrene, polyethylene, polypropylene, polyester, polyacrylonitrile, fluororesin, agarose, cellulose, nitrocellulose, dextran, sephadex, sepharose, liposome, carboxymethyl cellulose, polyacrylamide, Polysterene, companion rock, filter paper, ion exchange resin, plastic film, plastic tube, polyamine-methyl vinyl-ether-maleic acid copolymer, amino acid copolymer, ethylene-maleic acid copolymer, nylon, metal, glass, glass beads, or Magnetic particles and the like can be used.
  • polystyrene polyethylene, polypropylene, polyester, polyacrylonitrile, fluororesin, agarose, cellulose, nitrocellulose, dextran, sephadex, sepharose, liposome, carboxymethyl cellulose, polyacrylamide,
  • Cell culture plates, ELISA plates, tubes, and polymeric membranes may be used as other solid substrates.
  • the substrate may have any possible shape, for example spherical (bead), cylindrical (inside the test tube or well), planar (sheet, test strip), more preferably a multi-well plate (eg, 24 well) , 96 well, 192 well, 384 well, 576 well, etc.).
  • the purified DENV EDIII was coated with cobalt on the surface and fixed to the bead using Dynabeads (Invitrogen, Norway), a magnetic bead that binds to the His-tag of the protein.
  • the ssDNA library (500 pmole) dissolved in 100 uL of binding buffer was incubated at 90°C for 3 minutes, and then incubated at 4°C for 30 minutes. Then, it reacted for 1 hour while gently shaking with DENV EDIII previously immobilized on a magnetic bead. Then, the bead was washed twice with a binding buffer to remove ssDNA not binding to DENV EDIII bound to the bead.
  • the protein and the elution buffer (20 mM Tris, 50 mM NaCl, 5 mM KCl, 5 mM MgCl 2 , 0.01% tween 20, 300 mM immidazole, pH 8.0) to separate the protein and ssDNA from the magnetic bead and The bound ssDNA was eluted.
  • the eluted ssDNA was precipitated using ethanol, dissolved in 60 uL of distilled water, and then amplified by PCR using i-pfu polymerase (Intron Biotechnology, Korea) using Forward primer and Biotin-bound Reverse primer.
  • the amount of ssDNA and the concentration of DENV EDIII were carried out while decreasing for strict screening, and the DENV of the remaining ssDNA was measured by measuring the concentration of ssDNA eluted from repeated screening with a UV spectrometer (UV spectrometer, Biochrom Libra S22 spectromether). The screening process was performed while confirming the degree of binding with EDIII. The degree of bonding (%) is as shown in FIG. 1.
  • the 11th repeatedly selected ssDNA was amplified through PCR using unmodified Forward primer and Reverse primer, cloned into pENTR/TOPO vector (TOPO TA Cloning kit, Invitrogen, USA), and E. coli TOP10 cells (Invitrogen, USA). After the clone into which the ssDNA was inserted was purified using a miniprep kit (GeneAll, Korea), the base sequence was analyzed (COSMO Genetech, Korea). As a result, it was possible to analyze the sequence shown in Table 1 below.
  • the amount of 6-FAM-labeled ssDNA aptamer bound to magnetic beads with DENV EDIII fixed was separated by fluorescence measurement by separating only 6-FAM-labeled ssDNA bound to DENV EDIII using a magnet (1420 Victor multilabel counter, PerkinElmer, USA).
  • a magnet (1420 Victor multilabel counter, PerkinElmer, USA.
  • Example 2 In order to measure whether the DV4 aptamer selected through the SELEX (Systematic Evolution of Ligands by Exponential enrichment) technique of Example 1 binds to the actual DENV EDIII, the present inventors confirm the presence or absence of binding through the EMSA (Electrophoric Mobility Shift Assay) method. Did. Recombinant DENV EDIII 5 uM and aptamers DV1, DV2, DV3, and DV4 1 uM were incubated in a binding buffer of 20 uL at room temperature for 1 hour, and then run on 6% acrylamide gel. As a result of sybr green and coomassie blue staining, as shown in FIG. 3, it was confirmed that the developed DV4 aptamer binds to DENV EDIII better than other aptamers.
  • SELEX Systematic Evolution of Ligands by Exponential enrichment
  • Sequence list designation order SEQ ID NO: 1 ssDNA library CACCTAATACGACTCACTATAGCGGATCCGA-N40-CTGGCTCGAACAAGCTTGC SEQ ID NO: 2 Forward primer cacctaatac gactcactat agcgga SEQ ID NO: 3 Reverse primer gcaagcttgt tcgagccag SEQ ID NO: 4 DV4 Aptamer CACCTAATACGACTCACTATAGCGGATACGAGGGAGAGTCGGATGGGGTGGTTGTGTTAGGTGTAGTGGGTCTGGCTCGAACAAGCTTGC
  • the DNA aptamer that specifically binds to the dengue virus envelope protein domain III (DENV EDIII) of the present invention has a strong binding force and excellent specificity to the dengue virus surface protein, and is significantly remarkable compared to the diagnosis using existing antibodies. Since dengue virus can be diagnosed with increased accuracy, it is expected that it can be effectively used in various products for diagnosis, such as diagnostic kits and diagnostic compositions.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Hematology (AREA)
  • Zoology (AREA)
  • Urology & Nephrology (AREA)
  • Organic Chemistry (AREA)
  • Virology (AREA)
  • Wood Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Pathology (AREA)
  • Biophysics (AREA)
  • General Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Analytical Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne un aptamère d'ADN se liant spécifiquement au domaine III de la protéine d'enveloppe du virus de la dengue (DENV EDIII), une composition le comprenant pour diagnostiquer la dengue, et un kit pour le diagnostic de la dengue. La recherche Intensive et approfondie, réalisée par les présents inventeurs, dans le développement d'un nouveau biomarqueur pour Le diagnostic de la dengue a conduit à la découverte d'un aptamère d'ADN capable de se lier spécifiquement au domaine III de la protéine d'enveloppe du virus de la dengue (DENV EDIII) présentant une forte affinité de liaison et une excellente spécificité pour DENV EDIII. L'aptamère d'ADN est supposé être plus stable que les procédés ELISA classiques utilisant des anticorps et, en tant que tel, peut être avantageusement utilisé dans le développement d'une composition pour le diagnostic de la dengue et d'un kit pour le diagnostic de la dengue.
PCT/KR2019/017998 2018-12-19 2019-12-18 Aptamère d'adn se liant spécifiquement à ediii et utilisation correspondante WO2020130626A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201980084587.XA CN113227378B (zh) 2018-12-19 2019-12-18 与登革病毒ediii特异性结合的dna适配体及其用途

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020180165204A KR102048812B1 (ko) 2018-12-19 2018-12-19 뎅기 바이러스 ediii에 특이적으로 결합하는 dna 압타머 및 이의 용도
KR10-2018-0165204 2018-12-19

Publications (1)

Publication Number Publication Date
WO2020130626A1 true WO2020130626A1 (fr) 2020-06-25

Family

ID=68731418

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2019/017998 WO2020130626A1 (fr) 2018-12-19 2019-12-18 Aptamère d'adn se liant spécifiquement à ediii et utilisation correspondante

Country Status (3)

Country Link
KR (1) KR102048812B1 (fr)
CN (1) CN113227378B (fr)
WO (1) WO2020130626A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102048812B1 (ko) * 2018-12-19 2019-11-26 주식회사 엠디헬스케어 뎅기 바이러스 ediii에 특이적으로 결합하는 dna 압타머 및 이의 용도
KR20230107461A (ko) 2022-01-08 2023-07-17 광운대학교 산학협력단 중증 뎅기열 면역진단기기를 이용한 중증 뎅기열 관제 시스템 및 이에 의한 중증 뎅기열 관제방법

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20170085150A (ko) * 2016-01-08 2017-07-24 전북대학교산학협력단 뎅기 바이러스 4가지 혈청형의 외막 도메인 ⅲ에 대한 단일클론 항체 및 이의 용도
US20170226511A1 (en) * 2013-11-13 2017-08-10 National University Of Singapore Aptamers for binding flavivirus proteins
KR102048812B1 (ko) * 2018-12-19 2019-11-26 주식회사 엠디헬스케어 뎅기 바이러스 ediii에 특이적으로 결합하는 dna 압타머 및 이의 용도

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9073981B2 (en) * 2008-10-13 2015-07-07 Institute For Research In Biomedicine Dengue virus neutralizing antibodies and use thereof
EP2532748B1 (fr) * 2011-06-07 2014-10-08 Postech Academy-industry Foundation Liaison spécifique d'aptamère d'ADN sur la troponine I cardiaque humaine
KR101520084B1 (ko) 2014-01-09 2015-05-29 주식회사 젠바디 뎅기열 바이러스의 표피단백질 도메인1에 특이적인 단클론 항체를 포함하는 뎅기열 바이러스 항체의 신속진단키트 및 그 제조 방법
US10551381B2 (en) * 2016-02-11 2020-02-04 Massachusetts Institute Of Technology Anti-dengue virus NS1 protein monoclonal antibodies
KR101837856B1 (ko) * 2016-10-18 2018-03-13 단국대학교 산학협력단 뎅기바이러스 혈청형 2 및 3의 메틸트랜스퍼라제 활성을 억제하는 rna 앱타머
KR101837855B1 (ko) * 2016-10-18 2018-03-13 단국대학교 산학협력단 뎅기바이러스 혈청형 2의 메틸트랜스퍼라제 활성을 억제하는 rna 앱타머
KR101923197B1 (ko) * 2017-09-25 2018-11-28 주식회사 엠디엡투스 Esat6에 특이적으로 결합하는 dna 압타머 및 이의 용도
KR101923198B1 (ko) * 2017-09-25 2018-11-28 주식회사 엠디엡투스 Tb7.7에 특이적으로 결합하는 dna 압타머 및 이의 용도
KR102016668B1 (ko) * 2018-12-19 2019-08-30 주식회사 엠디헬스케어 치쿤군야 바이러스 e2에 특이적으로 결합하는 dna 압타머 및 이의 용도
KR101970963B1 (ko) * 2018-12-19 2019-04-23 주식회사 엠디헬스케어 황열 바이러스 ediii에 특이적으로 결합하는 dna 압타머 및 이의 용도

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170226511A1 (en) * 2013-11-13 2017-08-10 National University Of Singapore Aptamers for binding flavivirus proteins
KR20170085150A (ko) * 2016-01-08 2017-07-24 전북대학교산학협력단 뎅기 바이러스 4가지 혈청형의 외막 도메인 ⅲ에 대한 단일클론 항체 및 이의 용도
KR102048812B1 (ko) * 2018-12-19 2019-11-26 주식회사 엠디헬스케어 뎅기 바이러스 ediii에 특이적으로 결합하는 dna 압타머 및 이의 용도

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHEN , HENG-LI ET AL.: "Selection and characterization of DNA aptamers targeting all four serotypes of dengue viruses", PLOS ONE, vol. 10, no. 6 , e013 1240, 2015, pages 1 - 13, XP055577078 *
GANDHAM, SAI HARI A. ET AL.: "Thioaptamers targeting dengue virus type-2 envelope protein domain III", BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, vol. 453, 2014, pages 309 - 315, XP029117093, DOI: 10.1016/j.bbrc.2014.09.053 *

Also Published As

Publication number Publication date
KR102048812B1 (ko) 2019-11-26
CN113227378B (zh) 2023-11-24
CN113227378A (zh) 2021-08-06

Similar Documents

Publication Publication Date Title
CN111693712B (zh) 一种采用核酸适配体检测新冠病毒SARS-CoV-2 N蛋白的方法
US9562907B2 (en) Aptamers screening method based on graphene without target immobilization and the aptamers obtained from the method
WO2020130624A1 (fr) Aptamère d'adn se liant spécifiquement à la protéine 2 d'enveloppe du virus chikungunya et utilisation correspondante
ES2617226T3 (es) Ensayos multiplexados basados en aptámeros
US20230228751A1 (en) Aptamers against sars-cov-2
CN111304208B (zh) 一种特异性结合p-糖蛋白的核酸适配体、制备方法及其应用
WO2020130627A1 (fr) Aptamère d'adn se liant de manière spécifique à ediii du virus de la fièvre jaune et utilisation correspondante
WO2020130626A1 (fr) Aptamère d'adn se liant spécifiquement à ediii et utilisation correspondante
CN114127282A (zh) 适体的筛选方法和使用适体的免疫分析方法
US10751359B2 (en) Nucleic acid aptamer specifically binding to avian influenza virus subtype H5 and method of detecting avian ifluenza virus subtype H5 using the same
CN109837281B (zh) 特异性结合s100p蛋白的核酸适配体及其筛选、鉴定和应用
KR20120077154A (ko) 카나마이신에 특이적으로 결합하는 ｄｎａ 앱타머
CN109444414A (zh) 应用AlphaLISA检测登革病毒的方法及对应的组合物、试剂盒
CN101523216A (zh) 人类免疫缺陷病毒系列蛋白抗体测定方法
CN114457083B (zh) 一组特异识别孔雀石绿的单链dna核酸适配体及其应用
KR20130032128A (ko) 돼지 생식기 호흡기 증후군 바이러스 특이적인 ｄｎａ 앱타머
JP2008175750A (ja) 被検物質の検出方法
US20230417749A1 (en) Polynucleotide nanostructures for detecting viral infections and other diseases
CN114410641B (zh) 一种用于检测风疹病毒的核酸适配体、试剂盒及应用
JP2010540921A (ja) ヒトｇｓｔｔ１に対する抗体（抗−ｈｇｓｔｔ１）の検出のための免疫学的分析方法
KR101480495B1 (ko) C형 간염바이러스의 표면 단백질 e2에 대한 압타머 및 이의 용도
Yu Rapid and Sensitive Detection of Escherichia coli O157: H7 Using a QCM Sensor based on Aptamers Selected by Whole-Bacterium SELEX and a Multivalent Aptamer System
CN113252892A (zh) 一种用于提高病原微生物抗原检测灵敏度的探针及试剂盒
US9353421B2 (en) Aptamers that are specific for immunoglobulin-binding cell wall proteins
KR101263449B1 (ko) 설파디메톡신에 특이적으로 결합하는 ｄｎａ 앱타머

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19899091

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19899091

Country of ref document: EP

Kind code of ref document: A1